Nikon TIRF Instructions Manual
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Nikon TIRF Instructions Manual

Nikon TIRF Instructions Manual

System for te2000
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M301 E
03.6.CF.2
TIRF SYSTEM FOR TE2000
< INSTRUCTIONS >

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Summary of Contents for Nikon TIRF

  • Page 1 M301 E 03.6.CF.2 TIRF SYSTEM FOR TE2000 < INSTRUCTIONS >...

  • Page 3: Introduction

    Safety Precautions (Read the following before use!) WARNING and CAUTION Symbols Although Nikon products are designed to provide the utmost safety during use, incorrect usage or failure to follow the safety instructions provided may cause personal injury or property damage. To ensure correct use, read the instruction manual carefully and thoroughly before using the instrument.
  • Page 4 Safety Precautions (Read the following before use!) 1. Intended product use / Verwendungszweck This system should be used only for microscopic observations. Do not use for any other purpose. Bitte verwenden Sie dieses System nur zu mikroskopischen Beobachtungen. Nicht für andere Zwecke verwenden.
  • Page 5 Safety Precautions (Read the following before use!) 4. Laser emission position The laser product label, laser warning label and the aperture label displayed on the microscope stage warn that laser beams are emitted from these areas. Never attempt to look into the objective or the areas near the objective from above or below the stage as the lens emits a powerful laser beam toward the top and bottom sides of the stage.
  • Page 6 Kassettenhalterung oder der Befestigungslöcher des Lampengehäuses austritt. 3. Do not remove the TIRF attachment / Die TIRF Beleuchtungsvorrichtung Do not remove the TIRF attachment from the microscope. Removing the attachment may cause the laser beam to be emitted through the mounting hole.
  • Page 7 If you discover that the system is not functioning properly, immediately stop use, switch off the power switches to all pieces of equipment, disconnect the power cables from the outlets, and contact your nearest Nikon representative. Auseinanderbauen kann zu elektrischen Schlägen oder Störungen führen. Keines der Teile dieses Systems darf auseinandergebaut werden.
  • Page 8 If water enters a component, immediately suspend use of the system, disconnect the power cord from the outlet, and contact your nearest Nikon representative. - 6 -...
  • Page 9 • Make sure that the system is not subject to strong vibrations, which may degrade image quality. Consult with your Nikon representative about preventive measures you may take when your system is installed.

  • Page 10: Table Of Contents

    Safety Precautions (Read the following before use!)................ 1 Name and Function of Each Part ..................... 9 1 System configuration..................... 9 2 Optical path diagram (TIRF microscopy) ..............10 3 TIRF attachment ......................11 4 TIRF cassette holder ....................15 5 Laser unit ........................16 6 Shutter control unit ......................

  • Page 11: Name And Function Of Each Part

    I Name and Function of Each Part System configuration Name and Function of Each Part System configuration (3) Optical-fiber cable (4) Two-laser (4) Three-laser unit unit (2) TIRF attachment (17) Epi-fl attachment (8) Solid-state (8) Solid-state laser adapter laser adapter 161C 161C (5) Air-cooled argon laser...

  • Page 12: Optical Path Diagram (Tirf Microscopy)

    Name and Function of Each Part Optical path diagram (TIRF microscopy) Optical path diagram (TIRF microscopy) (11) Specimen (10) Objective TIRF attachment TIRF cassette holder (9) Dichroic mirror Detach the excitation filter. (7) ND filter (ND16) (8) Condenser lens (6) ND filter (ND2, ND8, ND32)

  • Page 13: Tirf Attachment

    Excitation light from a laser unit is guided into this unit through an optical-fiber cable. Excitation light emitted by this unit forms an image on the pupil surface of the objective. To protect your eyes and skin from exposure to the laser, do not detach the TIRF attachment from the microscope.
  • Page 14 The minimum diameter of the field diaphragm opening is approximately 40 µm at the specimen when using a 100x objective. The maximum area that can be illuminated with TIRF illumination varies from one objective to another. • When PlanAPO100x/1.45 is used: Approx. 110 µm at the specimen •...
  • Page 15 Indices (circumferential grooves) are provided to indicate various positions of the focal point adjustment knob that correspond to the objectives used for TIRF observation. Use these indices to guide laser focal point adjustment. Never attempt to look into the laser beam. (FDA CDRH: Class II, EN60825-1: Class 2, IEC60825-1: Class 2) The objective emits a powerful laser beam.
  • Page 16 Name and Function of Each Part TIRF attachment (6) Specimen cover Illustration of the specimen cover (6) Specimen cover This cover employs a filter that can attenuate the intensity of light in the proximity of 488 nm and 523 nm wavelengths by a factor of up to 1000. Make sure that the cover is always in place, except when you are adjusting the laser focal point.

  • Page 17: Tirf Cassette Holder

    TIRF cassette holder Filter blocks that incorporate dichroic mirrors are attached. This holder reflects the laser beam from the TIRF attachment to guide it to the objective. Up to six filter blocks can be attached and replaced as necessary. To protect your eyes and skin from exposure to the laser, do not detach the TIRF cassette holder from the microscope.

  • Page 18: Laser Unit

    Never touch the air-cooled argon, He-Ne laser, Solid-State laser, or excitation light optical-fiber cable. Contact your nearest Nikon representative if you find it necessary to detach the laser head from the laser unit due to laser failure or other problems.
  • Page 19 I Name and Function of Each Part Laser unit (1) Air-cooled argon laser (2) He-Ne laser (2) He-Ne laser (3) He-Ne laser, (3) He-Ne laser, Solid-State laser Solid-State laser (6) Caution (4) ND filter slider label Use this slider to change laser emission intensity.

  • Page 20: Shutter Control Unit

    Name and Function of Each Part Shutter control unit Shutter control unit This unit is designed to open or close the motorized shutter housed within each laser unit. Although the unit can open or close up to three channels of motorized shutters, for safety reasons, operation of motorized shutters is conducted through mutually exclusive control, as a result of which only one shutter can be opened at a time.
  • Page 21 I Name and Function of Each Part Shutter control unit (5) Eyepiece shutter closed lamp Lights yellow when the eyepiece shutter is closed. The motorized shutters are all closed when the lamp is not lit (and the eyepiece shutter is open).

  • Page 22: Eyepiece Shutter

    TIRF observation, while the mechanism closes the shutter in the laser unit during normal observation (observation using eyepieces) so that the laser beam is not guided into the TIRF attachment. An illumination pillar switch to be mounted on the microscope's illumination pillar is supplied with the eyepiece shutter.

  • Page 23: Operations

    II Operations Getting started (Emitting the laser) Operations Items to check before turning on the laser Before turning on the laser, check that 1) an objective or cap mounted on each of the sockets on the nosepiece of the microscope; 2) that the diascopic illumination lamphouse is attached;...

  • Page 24: Getting Started (Emitting The Laser)

    Operations Getting started (Emitting the laser) Getting started (Emitting the laser) (1) Switch on the power switch Emitting the argon laser 1 Switch on the power switch and turn the key switch clockwise on the argon laser power unit to turn on the unit. (2) Turn the key switch clockwise Argon laser power unit 2 Turn the key switch clockwise on the argon...
  • Page 25 II Operations Getting started (Emitting the laser) Emitting the He-Ne laser Turn the key switch on the He-Ne laser power unit clockwise to turn on the unit. Turn the key switch clockwise With the three-laser unit, another key switch is provided on the unit; this key switch is also turned clockwise.

  • Page 26: Operations (Tirf Microscopy)

    Achieving focus with epi-fl microscopy Use the epi-fl attachment to bring the specimen into focus, then use the TIRF attachment for microscopy. Open the eyepiece shutter. (All motorized shutters in the laser units will close, and the laser beam is interrupted.) Switch the microscope's optical path to the binocular section.

  • Page 27: Turning Off Power

    Rotate the excitation method switching ring of the epi-fl cassette holder to select a hollow without the filter block. Rotate the excitation method switching ring of the TIRF cassette holder to switch to a desired excitation method. Close the eyepiece shutter and switch the microscope's optical path to a port to which the TV camera is attached.

  • Page 28: Simple Surface Reflection Interference Observations Using Epi-Fl Attachment

    In theory, it is possible to make high-contrast observations of areas in contact with the cover glass using TIRF illumination. In practice, it is difficult to determine from images using TIRF illumination alone whether specimens such as cell cultures are actually in contact with the cover glass.

  • Page 29: Maintenance

    III Maintenance Replacing the halogen lamp Maintenance Replacing the halogen lamp If the halogen lamp for diascopic illumination has burned out, first check that all lasers are off, then follow the steps given below to replace the halogen lamp. Do not remove the lamphouse when the laser is on. Never attempt to remove the diascopic illumination lamphouse when the laser is on.

  • Page 30: Replacing The Objective And The Filter Block

    • Detach the laser from the laser unit. • Detach the optical-fiber cable from the laser unit. • Detach the optical-fiber cable from the TIRF attachment. • Detach the TIRF attachment from the microscope. Cleaning the lens section When cleaning optical components, use a soft, clean cotton cloth, lens tissue, or gauze lightly dampened with absolute alcohol (ethyl or methyl alcohol) to gently wipe off dirt or marks.

  • Page 31: Shutter Control Unit Communications Commands

    IV Shutter Control Unit Communications Commands Communication-controlled operation Shutter Control Unit Communications Commands Communication-controlled operation You can control this system from your PC or other devices through the RS-232C interface. Create your own control program based on the information given in this manual. 1-1 Communications system Start-stop serial communication 1-2 Connector specification...

  • Page 32: Cable Specification

    Shutter Control Unit Communications Commands Communication-controlled operation 1-3 Cable specification * To ensure trouble-free operation, use the supplied RS-232C cable for connecting your PC. 1-4 Communications parameters Baud rate: 9600 bps (fixed) Data length: 8 bits (fixed) Start bits: 1 bit (fixed) Stop bits: 1 bit (fixed) Parity check: None...

  • Page 33: Communications Sequence

    (five commands) Processing inhibit Processing is inhibited error Processing stop Command processing is stopped error 1-6 Communications sequence [PC] [TIRF system] c command Command o or n response [PC] [TIRF system] r command Command a or n response - 31 -...

  • Page 34: Commands

    Shutter Control Unit Communications Commands Communication-controlled operation 1-7 Commands (1) c command: Operation, control, and data setting command o: Normal response c command Description of operation n: Error response Opens a specified shutter and closes any other shutters. If an open time period has been specified, the shutter will close when the specified period elapses.

  • Page 35: Troubleshooting

    Laser beam focal point is near Adjust the laser position. the center of the objective's pupil. TIRF image with poor contrast The laser beam emitted by the Adjust the position of the condenser objective is not a parallel ray.

  • Page 36: Performance And Specifications

    Performance and Specifications Performance and Specifications System configuration Inverted microscope TE2000-E/U/S, eyepiece shutter, TIRF attachment, shutter control unit, RS-232C cable, AC adapter, laser unit (two-laser unit or three-laser unit), optical-fiber cable. Applicable objective PlanApo TIRF 100×/1.45 PlanApo TIRF 60×/1.45 PlanApo 60×/1.40 Applicable Lasers •...
  • Page 37 VI Performance and Specifications AC adapter for the Manufacturer: PHIHONG ENTERPRISE (Taiwan) shutter control unit Model: PSA-30U-120 (N) Rated input voltage: AC 100-240 V, 0.7 A, 50/60 Hz Rated output voltage: DC 12 V Rated output current: 2.5 A Electric shock prevention class: Class I Others: UL-recognized product, GS-approved product，...

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